Difference between revisions of "Part:BBa K892403"

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===Usage and Biology===
 
===Usage and Biology===
This part was tested by the [http://2012.igem.org/Team:Washington 2012 University of Washington] iGEM team to bind to various subtypes of Hemagglutinin in Group 1.To test BBa _K892401, it was inserted into a protein expression vector, pETCON. HB36.5 was then produced and transformed into yeast as described in the [http://2012.igem.org/Team:Washington/Protocols/Yeast  2012 Team's Yeast Transformation Protocol]. The binding protein was then tested for activity against H2. For a detailed description of the methodology, please see the [http://2012.igem.org/Team:Washington/Protocols/Display 2012 UW iGEM Yeast Surface Display protocol]. The resulting data is shown below.
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This part was tested by the [http://2012.igem.org/Team:Washington 2012 University of Washington] iGEM team to bind to various subtypes of Hemagglutinin in Group 1.To test BBa _K892401, it was inserted into a protein expression vector, pETCON. HB36.5_H312A was then produced and transformed into yeast as described in the [http://2012.igem.org/Team:Washington/Protocols/Yeast  2012 Team's Yeast Transformation Protocol]. The binding protein was then tested for activity against H2. For a detailed description of the methodology, please see the [http://2012.igem.org/Team:Washington/Protocols/Display 2012 UW iGEM Yeast Surface Display protocol]. The resulting data is shown below.
  
[[Image:UWashington FLU HB36 results graph.png|border|800px|center|thumb|HB36.5 and its variants tested against hemagglutinin subtype 2. HB36.5 is shown to have very little activity in binding H2.]]
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[[Image:UWashington FLU HB36 results graph.png|border|800px|center|thumb|HB36.5 and its variants tested against hemagglutinin subtype 2. HB36.5 is shown to have very little activity in binding H2. The single-point mutant H312A dramatically increased binding to H2 at all concentrations of HA tested.]]
  
  

Revision as of 16:24, 3 October 2012

HB36.5_H312A

A mutated HB80.4 flu binder aimed to increase binding to H2, a subtype of hemagglutinin on the surface of the Influenza virus. This mutant has a point mutation at residue 312 from Histidine to Alanine, which dramatically increases the binding to H2 over the native HB36.5 binder.

Usage and Biology

This part was tested by the [http://2012.igem.org/Team:Washington 2012 University of Washington] iGEM team to bind to various subtypes of Hemagglutinin in Group 1.To test BBa _K892401, it was inserted into a protein expression vector, pETCON. HB36.5_H312A was then produced and transformed into yeast as described in the [http://2012.igem.org/Team:Washington/Protocols/Yeast 2012 Team's Yeast Transformation Protocol]. The binding protein was then tested for activity against H2. For a detailed description of the methodology, please see the [http://2012.igem.org/Team:Washington/Protocols/Display 2012 UW iGEM Yeast Surface Display protocol]. The resulting data is shown below.

HB36.5 and its variants tested against hemagglutinin subtype 2. HB36.5 is shown to have very little activity in binding H2. The single-point mutant H312A dramatically increased binding to H2 at all concentrations of HA tested.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 107
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 107
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 289
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 107
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 107
  • 1000
    COMPATIBLE WITH RFC[1000]